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背景:凋亡参与了脑出血后的神经损伤机制,凋亡是一个需要能量的过程。但目前尚缺乏足够的直接证据证实脑出血后能量供应变化与凋亡发生的相关性。目的:研究脑出血后脑组织线粒体ATPase-6基因表达变化与细胞凋亡的相关性。设计:完全随机对照实验。地点和对象:实验在解放军第三军医大学野战外科研究所神经内科实验室完成,对象为Wister大鼠40只干预:以50μL鼠尾血注入大鼠尾状核制作大鼠脑出血模型,注血后分别于12h,1,3,7d处死大鼠,断头取血肿周围脑组织;对照组仅进针不注血。用RT-PCR方法测定以上不同时相点大鼠脑出血后血肿周围水肿组织线粒体ATPase-6基因表达情况。TdT介导的dUTP缺口末端标记法检测TUNEL阳性细胞的表达。主要观察指标:各组大鼠线粒体ATPase-6基因表达,TUNEL阳性细胞表达。结果:出血后12h,1,3d线粒体ATPase-6基因表达较对照组下降,但差异不显著(P>0.05)。至第7天时表达明显下降(P<0.01)。TUNEL阳性细胞在出血后12h开始表达犤(24.33±2.50)个犦,3d达到高峰犤(214.17±21.45)个,7d时仍持续较高水平(124.88±18.94)个犦。结论:脑出血早期血肿周围水肿组织线粒体ATPase-6基因表达无明显减少,晚期表达明显下降,在出血早期存在凋亡发生的能量基础。
BACKGROUND: Apoptosis is involved in the mechanism of neural damage after intracerebral hemorrhage. Apoptosis is a process that requires energy. However, there is not enough direct evidence to confirm the correlation between energy supply and apoptosis after intracerebral hemorrhage. Objective: To study the correlation between the change of mitochondrial ATPase-6 gene expression and apoptosis after intracerebral hemorrhage in rats. Design: Complete randomized controlled experiment. Location and Subjects: The experiment was performed in the Neurology Laboratory of the Institute of Field Surgery, Third Military Medical University of Chinese PLA. Subjects were 40 Wistar rats. Intervention: Wistar rats were injected with 50 microliters rat caudate nucleus into caudate to produce rat intracerebral hemorrhage model. After 12h, 1,3,7d, respectively, rats were sacrificed, decapitated to take the brain tissue around the hematoma; the control group only needle injection without blood. The expression of mitochondrial ATPase-6 gene in edema around the hematoma after intracerebral hemorrhage at different time points was measured by RT-PCR. TdT-mediated dUTP nick end labeling was used to detect TUNEL positive cells. MAIN OUTCOME MEASURES: Mitochondrial ATPase-6 gene expression and TUNEL positive cells were observed in all groups. Results: The mitochondrial ATPase-6 gene expression at 12h, 1,3d after hemorrhage was lower than that in control group, but the difference was not significant (P> 0.05). The expression was significantly decreased on the 7th day (P <0.01). TUNEL positive cells expressed (.3) (24.33 ± 2.50) 12 at 12h after hemorrhage, peaked at 3d 3d (214.17 ± 21.45), and persisted to a high level (124.88 ± 18.94) 7 at 7d. CONCLUSION: The expression of mitochondrial ATPase-6 gene in edema tissue around the hematoma in the early stage of ICH has no obvious decrease and the late phase expression is obviously decreased. The energy base of apoptosis appears in the early stage of hemorrhage.